Xylan degradation improved by a combination of monolithic columns bearing immobilized recombinant β‐xylosidase from Aspergillus awamori X‐100 and Grindamyl H121 β‐xylanase

Abstract

Synergistic action of exo- and endohydrolazes is preferred for effective destruction of biopolymers. The main purpose of the present work was to develop an efficient tool for degradation of xylan. Macroporous lab-made monolithic columns and commercial CIM-Epoxy disk were used to immobilize the recombinant β-xylosidase from Aspergillus awamori and Grindamyl β-xylanase. The efficiency of xylan degradation using the low-loaded β-xylosidase column appeared to be four times higher than for the in-solution process and about six times higher than for the high-loaded bioreactor. Disk bioreactor with the Grindamil β-xylanase operated in a recirculation mode has shown noticeable advantages over the column design. Additionally, a system comprised of two immobilized enzyme reactors (IMERs) was tested to accelerate the biopolymer hydrolysis, yielding total xylan conversion into xylose within 20 min. Fast online monitoring HPLC procedure was developed where an analytical DEAE CIM disk was added to the two-enzyme system in a conjoint mode. A loss of activity of immobilized enzymes did not exceed 7% after 5 months of the bioreactor usage. We can therefore conclude that the bioreactors developed exhibit high efficiency and remarkable long-term stability.

Efficient and rapid digestion of proteins with a dual-enzyme microreactor featuring 3-D pores formed by dopamine/polyethyleneimine/acrylamide-coated KIT-6 molecular sieve

“…PEI, a branched polymer copolymer with abundant amino groups, readily links enzymes using glutaraldehyde (GA) as a spacer. The chain structure of PEI and GA extends the connected enzyme, reducing spatial hindrance 26 . Additionally, residual double bonds react with thiol groups on proteins, facilitating rapid and gentle immobilization of another enzyme without affecting the activity of the first immobilized enzyme 27 , 28 .…”

Immobilized enzyme reactors based on monoliths: Effect of pore size and enzyme loading on biocatalytic process

“…Moreover, the advantages of methacrylate‐based monolithic matrices include mechanical and chemical stability, as well as the possibility of synthesis of these materials in various geometrical formats, namely, in the shape of disks, columns or tubes . Owing to the mentioned advantages of monoliths, they have become popular not only for application in different kinds of chromatography and solid‐phase extraction , but also for the preparation of flow‐through immobilized enzyme reactors (IMERs) suitable for proteomics , microfluidic analysis , pharmaceutics , biodiesel and oligosaccharides production, etc. In most cases, the covalent binding of enzyme to the surface of macroporous monoliths was applied .…”